Research methods assessing sodium hypochlorite cytotoxicity: A scoping review

Sodium hypochlorite (NaOCl) cytotoxicity has been assessed using different methodologies, which has led to arbitrary interpretations. This scoping review aimed to discuss the different methodological protocols for assessing NaOCl cytotoxicity. A literature review performed in the PubMed and Embase databases up to July 2023 identified manuscripts reporting NaOCl cytotoxicity. The dataset identified 546 publications, further screened by 2 reviewers. Ninety studies were identified and mined methodologically to collect information on cell type, cytotoxicity assay, NaOCl dilution solutions, presence of fetal bovine serum (FBS), and NaOCl exposure time. The culture medium used in cytotoxicity assays contains buffering substances that neutralize the pH of NaOCl, thus reducing its cytotoxicity, an approach that may lead to bias when solutions with different pH are compared. For short exposure periods, as in simulations to evaluate the contact between irrigant and periapical tissue cells during chemo-mechanical preparation, NaOCl dilution should be performed with saline, which does not buffer the irrigant. For long exposure periods, as in simulations of irrigant extrusions, NaOCl should be diluted in the culture medium, to reproduce the expected buffering effect occurring in extrusions. The presence of FBS in culture medium can decrease NaOCl toxicity. There is no standardization of NaOCl cytotoxicity methodologies. This poses the risk of arriving at incorrect results and, therefore, pertinent tests must be refined.